epa.gov Regulations and Standards October 1980 Washington DC ...Nitrobenzene, also known as...
Transcript of epa.gov Regulations and Standards October 1980 Washington DC ...Nitrobenzene, also known as...
AmbientWater QualityCriteria forNitrobenzene
;EPA
United StatesEnvironmental ProtectionAgency
Office of WaterRegulations and StandardsCriteria and Standards DivisionWashington DC 20460
EPA 440/5-80-061October 1980
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AMBIENT WATER QUALITY CRITERIA FOR
NITROBENZENE
Prepared ByU.S. ENVIRONMENTAL PROTECTION AGENCY
Office of Water Regulations and StandardsCriteria and Standards Division
Washington, D.C.
Office of Research and DevelopmentEnvironmental Criteria and Assessment Office
Cincinnati, Ohio
Carcinogen Assessment GroupWashington, D.C.
Environmental Research LaboratoriesCorvalis, OregonDuluth, Minnesota
Gulf Breeze, FloridaNarragansett, Rhode Island
DISCLAIMER
This report has been reviewed by the Environmental Criteria and
Assessment Office, U.S. Environmental Protection Agency, and approved
for publication. Mention of trade names or commercial products does not
constitute endorsement or recommendation for use.
AVAILABILITY NOTICE
This document is available to the public through the National
Technical Information Service, (NTIS), Springfield, Virginia 22161.
FOREWORD
Section 304 (a)(I) of the Clean Water Act of 1977 (P.L. 95-217),requires the Administrator of the Environmental Protection Agency topub 1i sh criteri a for water qual ity accurately reflecting the 1atestscientific knowledge on the kind and extent of all identifiable effectson hea1th and we 1fare wh ich may be expected from the presence ofpollutants in any body of water, including ground water. Proposed waterquality criteria for the 65 toxic pollutants listed under section 307(a)(1) of the Cl ean Water Act were developed and a not ice of thei ravailability was published for public comment on March 15, 1979 (44 FR15926), July 25, 1979 (44 FR 43660), and October 1, 1979 (44 FR 56628).This document is a revision of those proposed criteria based upon aconsideration of comments received from other Federal Agencies, Stateagencies, special interest groups, and individual scientists. Thecriteria contained in this document replace any previously published EPAcriteria for the 65 pollutants. This criterion document is alsopublished in satisifaction of paragraph 11 of the Settlement Agreementin Natural Resources Defense Council, et. al. vs. Train, 8 ERC 2120(D.D.C. 1976), modified, 12 ERC 1833 (D.D.C. 1979).
The term "water quality criteria" is used in two sections of theClean Water Act, section 304 (a) (1) and section 303 (c)(2). The term hasa different program impact in each section. In section 304, the termrepresents a non-regulatory, scientific assessment of ecological effects. The criteria presented in this publication are such scientificassessments. Such water quality criteria associated with specificstream uses when adopted as State water quality standards under section303 become enforceable maximum acceptable levels of a pollutant inambient waters. The water quality criteria adopted in the State waterquality standards could have the same numerical limits as the criteriadeveloped under section 304. However, in many situations States may wantto adjust water quality criteria developed under section 304 to reflectlocal environmental conditions and human exposure patterns beforeincorporation into water quality standards. It is not until theiradoption as part of the State water quality standards that the criteriabecome regulatory.
Guidelines to assist the States in the modification of criteriapresented in this document, in the development of water qualitystandards, and in other water-related programs of this Agency, are beingdeveloped by EPA.
STEVEN SCHATZOWDeputy Assistant AdministratorOffice of Water Regulations and Standards
iii
ACKNOWLEDGEMENTS
Aquatic Life Toxicology:
William A. Brungs, ERL-NarragansettU.S. Environmental Protection Agency
David J. Hansen, ERL-Gulf BreezeU.S. Environmental Protection Agency
Mammalian ToxicolQgy and Human Health Effects:
Karl Gabriel (author)Medical College of Pennsylvania
Steven D. Lutkenhoff (doc. mgr.)ECAO-CinU.S. Environmental Protection Agency
Si Duk Lee (doc. mgr.), ECAO-CinU.S. Environmental Protection Agency
Patrick DurkinSyracuse Research Corporation
Sherwin KevyChildren's Hospital Medical Center
David J. McKee, ECAO-RTPU.S. Environmental Protection Agency
Alan B. RubinU.S. Environmental Protection Agency
James WitheyHealth and Welfare, Canada
John AutianUniversity of Tennessee
J. P. Bercz, HERLU.S. Environmental Protection Agency
Richard CarchmanMedical College of Virginia
Thomas J. HaleyNational Center for Toxicological Res.
Van KozakUniversity of WiscQnsin
V.M. Sadagopa RamanujamUniversity of Texas Medical Branch
Carl SmithUniversity of Cincinnati
Technical Support Services Staff: D.J. Reisman, M.A. Garlough, B.L. Zwayer,P.A. Daunt, K.S. Edwards, T.A. Scandura, A.T. Pressley, C.A. Cooper,M.M. Denessen.
Clerical Staff: C.A. Haynes, S.J. Faehr, L.A. Wade, D. Jones, B.J. Bordicks,B.J. Quesnell, P. Gray, R. Rubinstein.
iv
TABLE OF CONTENTS
Criteria Summary
Introduction
Aquatic Life ToxicologyIntroductionEffects
Acute Toxi cityChronic ToxicityPlant EffectsSummary
CriteriaReferences
Mammalian Toxicology and Human Health EffectsIntroductionExposure
Ingestion from WaterIngestion from FoodInhalationDermal
PharmacokineticsAbsorptionDistributionMetabolismExcretion
EffectsAcute, Subacute, and Chronic ToxicitySynergism and/or AntagonismTeratogeni ci tyMutagenicityCarci nogen i city
Criteria FormulationExisting Guidelines and StandardsCurrent Levels of ExposureSpecial Groups at RiskBasis and Derivation of Criterion
References
Appendix
v
A-l
B-1B-1B-1B-1B-1B-2B-2B-2B-7
C-lC-lC-2C-2C-4C-5C-6C-8C-8C-9C-llC-14C-19C-19C-24C-24C-25C-25C-27C-27C-27C-28C-28C-3l
C-45
CRITERIA DOCUMENT
NITROBENZENE
CRITERIA
Aquatic Life
The available data for nitrobenzene indicate that acute toxicity to
freshwater aquatic life occurs at concentrations as low as 27,000 pg/l and
would occur at lower concentrations among species that are more sensitive
than those tested. No definitive data are available concerning the chronic
toxicity of nitrobenzene to sensitive freshwater aquatic life.
The available data for nitrobenzene indicate that acute toxicity to
saltwater aquatic life occurs at concentrations as low as 6,680 pg/1 and
would occur at lower concentrations among species that are more sensitive
than those tested. No definitive data are available concerning the chronic
toxicity of nitrobenzene to sensitive saltwater aquatic life.
Human Health
For compari son purposes, two approaches were used to deri ve cri teri on
levels for nitrobenzene. Based on available toxicity data, for the protec
tion of public health, the derived level is 19.8 mg/1. Using available
organoleptic data, for controlling undesirable taste and odor qualities of
ambient water, the estimated level is 30 pg/l. It should be recognized that
organoleptic data as a basis for establishing a water quality criterion have
limitations and have no demonstrated relationship to potential adverse human
health effects.
vi
INTRODUCTION
Nitrobenzene is produced for industrial use by the nitration of benzene
with nitric and sulfuric acids. Estimates of annual nitrobenzene production
range from 200 to over 700 million pounds (Dorigan and Hushon, 1976; Lu and
Metcalf, 1975). The principal use of nitrobenzene is for reduction to ani
line, which is widely used as an ingredient for dyes, rubber, and medicinals
(McGraw-Hill, 1971; Kirk and Othmer, 1967). The commercial applications of
nitrobenzene are: reduction to aniline (97 percent), solvent for Friedel
Crafts reaction, metal polishes, shoe black, perfume, dye intermediates,
crystallizing solvent for some substances, and as a combustible propellant
(Dorigan and Hushon, 1976).
Nitrobenzene is stored in closed containers and is not usually released
to the open air. Atmospher;'c contamination is usually prevented in plants
manufacturing or using nitrobenzene by the use of activated charcoal ab
sorbers or a carbon dioxide blanket. There is no industrial monitoring of
nitrobenzene in the atmosphere. The greatest loss of nitrobenzene during
production (estimated as eight million pounds annually) occurs at the acid
extraction step in the purification of the crude reaction mixture, when
nitrobenzene is lost to the effluent wash (Dorigan and Hushon, 1976). Thus,
the greatest exposure to nitrobenzene occurs inside plants and most cases of
chronic nitrobenzene exposure in man are nitrobenzene workers. Today plant
levels of nitrobenzene are usually kept below the threshold limit value
(TLV) of 5 mg/m3 [Goldstein, 1975; American Conference of Governmental
Industrial Hygienists (ACGIH), 1977] but much higher levels have been re
ported in the oast (Pacseri and Magos, 1958). Nitrobenzene may also form
spontaneous ly in the atmosphere from the photochemi ca 1 reacti on of benzene
with oxides of nitrogen.
A-1
Nitrobenzene, also known as nitrobenzo1, essence of mirbane, and oil of
mirbane, is a pale yellow oily liquid with an almond-like odor (Kirk and
Othmer, 1967). The color of the liquid varies from pale yellow to yellowish
brown depending on the purity of the compounds (Kirk and Othmer, 1967). In
the solid state it forms bright yellow crystals. Nitrobenzene,
C6H5N02, has a molecular weight of 123.11 g.
The physical properties of nitrobenzene are as follows: a boiling point
of 210° to 211°C at 760 mm Hg, a melting point of 6°C, a density of 1.205 at
15°C, a refractive index of 1.5529, and a flash point of 89°C (Stecher,
1968). It is steam volatile (Stecher, 1968) and at 25°C nitrobenzene has a
vapor pressure of 0.340 mm Hg (Jordan, 1954).
Ni trobenzene is mi sci b1e wi th most organi c solvents, such as ethano1,
diethy1 ether, acetone, and benzene (Kirk and Othmer, 1967). It is slightly
soluble in water, 0.1 per 100 parts of water (1,000 mgtl) at 20°C (Kirk and
Othmer, 1967). In aqueous solutions, nitrobenzene has a sweet taste (Kirk
and Othmer, 1967).
Nitrobenzene undergoes substitution reactions but requires more vigorous
conditions than does benzene. Substitution takes place at either the
meta-(3) position or the ortho-(2) or para-(4) positions depending on the
physical conditions (Kirk and Othmer, 1967). Nitrobenzene undergoes photo
reduction when irradiated with ultraviolet light in organic solvents that
contain abstractab1e hydrogen atoms (Bar1trop and Bunce, 1968).
Nitrobenzene is a fairly strong oxidizing agent (Kirk and Othmer, 1967;
Millar and Springfield, 1966). Since the compound can act as an oxidizing
agent in the presence of aqueous solutions of alkali hydroxides, it has the
capability of oxidizing compounds containing free phenolic hydroxyl groups
without effectively changing these groups (Millar and Springfield, 1966).
A-2
Nitrobenzene is rective and will undergo nitration, halogenation, and sulfo
nation by the same methods used for benzene. However, these reactions are
unlikely to occur in environmental conditions.
The reduction of nitrobenzene to aniline probably outranks all other
uses of nitrobenzene as an industrial chemical (Kirk and Othmer, 1967). The
di- and the trinitrobenzenes are used in military and industrial explosives.
A-3
REFERENCES
American Conference of Governmenta 1 Industri a1 Hygieni sts. 1977.
tation of the threshold limit value for substances in workroom air.
nati, Ohio.
Documen
Cincin-
Barltrop, A.J. and N.J. Bunce. 1968. Organic photochemistry, Part 4. The
photochemical reduction of nitro-compounds. Jour. Chem. Soc. Sec. C.
12: 1467.
Dorigan, J.
nitrobenzene.
and J. Hushon. 1976.
U.S. Environ. Prot. Agency.
Air pollution assessment of
Goldstein, 1. 1975. Studies on MAC values of nitro- and amino-derivatives
of aromatic hydrocarbons. Adverse Effects Environ. Chem. Psych. Drugs.
1: 153.
Jordan, T.E. 1954. Vapor Pressure of Organic Compounds. Interscience Pub
1i shers, Inc., New York.
Kirk, R.E. and D.F. Othmer (eds.) 1967. Kirk-Othmer Encyclopedia of Chemi
cal Technology. 2nd ed. John Wiley and Sons, Inc., New York.
Lu, P.Y. and R. Metcalf. 1975. Environmental fate and biodegradability of
benzene derivatives as studies in a model aquatic ecosystem. Environ.
Health Perspect. 19: 269.
A-4
McGraw-Hill. 1971. Encyclopedia of Science and Technology. McGraw-Hill
Book Co., New York.
Mi 11 ar, 1.T. and H. D. $pri ngfi e1d (eds.) 1966. $i dgwi ck I s Organi c Chemi s
try of Nitrogen. 3rd ed. Clarendon Press, Oxford.
Pacseri, 1. and L. Magos. 1958. Determination of the measure of exposure
to aromatic nitro and amino compounds. Jour. Hyg. Epidemiol. Microbiol.
Inrnuno1. 2: 92.
Stecher, P.G. (ed.) 1968. The Merck Index. 8th ed. Merck and Co., Inc.,
Rahway, New Jersey.
A-5
AQuatic Life Toxicology*
INTRODUCTI ON
Static tests with the bluegill~ Daphnia magna~ and the alga~ Selenastrum
capricornutum~ indicate little difference in sensitivity with no SO percent
effect concentration lower than 27~OOO ~g/l. An embryo-larval test with the
fathead minnow demonstrated no adverse effects at the highest test concen-
tration of 32~OOO ~g/l.
Static acute tests with the sheepshead minnow and Mysidopsis bahia indi
cate that the latter is much more sensitive to nitrobenzene. Adverse ef-
fects were observed on a saltwater alga at concentrations slightly higher
than the LCSO for the mysid shrimp.
EFFECTS
Acute Toxicity
The 48-hour EC SO for Daphnia magna and the 96-hour LC SO for the
bluegill are 27~000 and 42~600 ~g/l~ respectively (Table 1).
The saltwater species are comparable to the freshwater species in their
sensitivity to nitrobenzene. The mysid shrimp LCSO is 6~680 ~g/l (Table
1) and the LCSO for the sheepshead minnow is S8~600 ~g/l.
Chronic Toxicity
No adverse effects were observed during an embryo-larval test with the
fathead minnow at test concentrations of nitrobenzene as high as 32~OOO ~g/l
(Table 2).
*The reader ;s referred to the Guidelines for Deriving Water Quality Criteri a for the Protection of AQuati c Life and Its Uses in order to betterunderstand the following discussion and recommendation. The followingtables contain the appropriate data that were found in the literature~ andat the bottom of each table are calculations for deriving various measuresof toxicity as described in the Guidelines.
B-1
Plant Effects
The 96-hour ECSO values for reduction of cell numbers and inhibition
of chlorophyll 2. in the freshwater alga, Selenastrum capricornutum, are
42,800 and 44,100 ~g/l, respectively (Table 3).
The cell numbers of Skeletonema costatum were reduced by SO percent at a
concentration of 9,6S0 ~g!l (Table 3). Chlorophyll a was equally inhibited
at a concentration of 10,300 ~g/l.
Summary
The acute 50 percent effect 1eve1s of Daphni a magna and the b1uegi 11
were 27,000 and 42,600 lJg!l, respectively. No effects on fathead minnow
embryos or larvae were observed at concentrations as high as 32,000 lJg/l. A
freshwater alga was of similar sensitivity with an ECSO value for chloro
phyll 2. of 44,100 ~g/l.
Ninety-six-hour LC SO values were 6,680 and S8,600 lJg/1 for the mysid
shrimp and sheepshead minnow, respectively. The ECSO
for cell numbers of
a saltwater alga was 9,6S0 ~g/l.
CRITER IA
The available data for nitrobenzene indicate that acute toxicity to
freshwater aquatic life occurs at concentrations as low as 27,000 ~g/l and
would occur at lower concentrations among species that are more sensitive
than those tested. No definitive data are available concerning the chronic
toxicity of nitrobenzene to sensitive freshwater aquatic life.
The available data for nitrobenzene indicate that acute toxicity to
saltwater aquatic life occurs at concentrations as low as 6,680 ~g/1 and
would occur at lower concentrations among species that are more sensitive
than those tested. No data are avialable concerning the chronic toxicity of
nitrobenzene to sensitive saltwater aquatic life.
B-2
Table 1. Acute values for nitrobenzene (U.S. EPA. 1918)
Species Method-LC50/EC50
(psI I )SpecIes AcuteValue (psI!)
fRESHWATER SPECIES
Cladoceran,Daphnia magna
61 ueglll,Lepomls macrochlrus
S, U
S, U
27,000
42,600
27,000
42,600
SALTWATER SPECIES
tJ:II
w
Mysld shrlq>,Mysldopsls bahla
Sheepshead minnow,Cyprlnodon varlegatus
S, U
S, U
6,680
58,600
6,680
58,600
* S = static, U = unmeasured
No final Acute Values are calculable since the minimum data baserequirements are not met.
SpecI..
Tabl. 2. Chronic val .... for .Ifrobeftz... (U.S. EPA. 1978)
CbrGIIlcVal.(ps/I)
FRESHWATER SPECIES
Fathead Illinnow.Plmephales promela$
E-L
* E-l = embryo-larval
No acute-chronic ratio 1$ calculable.
I.bl. 3. PI.nt V11lu8$ for .Itrobenzen. (U.S. EPA, 1918)
Species Effect
FRESHWATER SPECIES
Resulthag/I)
Alga,Selenastrum caprlcornutum
Alga,Selenastrum caprlcornutum
96-hr EC50ch lorophy II .!.
96-hr EC50ce I I numbers
44,100
42,800
SALTWATER SPECIES
tXlI
U1
Alga,Skeletonema costatum
Alga,Skeletonema costatum
96-hr EC50cell numbers
96-hr EC50ch lorophy I I .!.
9,650
10,300
REFERENCES
u.S. EPA. 1978. In-depth studies on health and environmental impacts of
selected water pollutants. U.S. Environ. Prot. Agency, Contract No.
68 -01-4646 •
B-6
Mammalian Toxicology and Human Health Effects
INTRODUCTION
Nitrobenzene, a pale yellow liquid at room temperature with a character
istic bitter almond aroma, is also known as oil of mirbane, nitrobenzo1, and
artificial bitter almond oil. It is produced for industrial use by the ni
tration of benzene with nitric and sulfuric acids. Estimates of annual ni
trobenzene production range from 200 to over 700 million pounds (Dorigan and
Hushon, 1976; Lu and Metcalf, 1975). The principal use of nitrobenzene is
for reduction to aniline, which is widely used as an ingredient for dyes,
rubber, and medicinals. The commercial applications of nitrobenzene are:
reduction to aniline (97 percent), solvent for Friede1-Crafts reaction, me
tal polishes, shoe black, perfumes, dye intermediates, crystallizing sol
vent, and as a combustible propellant (Dorigan and Hushon, 1976).
Nitrobenzene is stored in closed containers and not usually released to
the open air. In plants manufacturing or using nitrobenzene, atmospheric
contamination is usually prevented by the use of activated charcoal absorb
ers or a carbon dioxide blanket. There is no industrial monitoring of ni
trobenzene in the atmosphere. The greatest loss of nitrobenzene during pro
duction (estimated as eight million pounds annually) occurs at the acid ex
traction step in the purification of the crude reaction mixture, when nitro
benzene is lost to the effluent wash (Dorigan and Hushon, 1976). Thus, the
greatest exposure to nitrobenzene occurs inside plants, while most cases of
chroni c ni trobenzene exposure inman i nvo 1ve nitrobenzene workers. Today,
plant levels of nitrobenzene are usually kept below the threshold limit
value (TLV) of 5 mg/m3 [Goldstein, 1975; American Conference of Governmen
tal Industrial Hygienists (ACGIH), 1977J but much higher levels have been
reported in the past (Pacseri and Magos, 1958). Nitrobenzene may also form
C-1
spontaneous ly in the atmosphere from the photochemi ca1 react ion of benzene
with oxides of nitrogen; the symptoms of nitrobenzene poisoning are similar
to the symptoms experienced by victims of Japanese photochemical smog (Dori
gan and Hushon, 1976).
Ni trobenzene can be detected for mon i tori ng purposes by co1orimetri c
reaction, or by collection on a charcoal filter, extraction, reduction to
an i1i ne, and product ion of a co1ored product by di azot izat ion of the an i
line. These methods can detect nitrobenzene from 1.0 to 500 mg/m3 (0.2 to
100 ppm) (Dorigan and Hushon, 1976). Nitrobenzene in wastewater can be mea
sured by gas chromatography (Austern, et ale 1975). Exposure of workers to
nitrobenzene is monitored by urinary level s of p-nitropheno1 (Piotrowski,
1967) and p-aminophenol (Pacseri and Magos, 1958).
Some of the physical and chemical properties of nitrobenzene are summar
ized in Table 1. Common derivatives of nitrobenzene (besides aniline) are
dinitrobenzene, nitrobenzene-sulfonic acid, and nitrochlorobenzene. There
are many other derivatives of nitrobenzene, and many of them are very hazar
dous to man as toxic agents, mutagens, and carcinogens.
EXPOSURE
Ingestion from Water
Nitrobenzene can be released into wastewater from production plants as
the result of losses during the production of nitrobenzene, aniline, or dye
stuffs. The solubility of nitrobenzene is low, and it produces a detectable
odor in water at a concentration as low as 0.03 mg/l (Austern, et ale 1975;
U.S. EPA, 1970; Alekseeva, 1964), so that large amounts can not readily ac
cumu1ate unnot iced. Leve1s of nitrobenzene in wastewater are mon i tored by
plants produc ing and us i ng the chemi ca1 but nitrobenzene 1eve1sin city
C-2
TABLE 1
Properties of Nitrobenzene
Formul a:
Molecular weight:
Freezing point:
Boiling point:
Water solubility:
Soluble in:
Vapor pressure:
Vapor density:
Log partition co-efficient:
Density:
Flash point:
Autoignition temp:
Viscosity:
Detection level of characteristic bitter almond odor:
*Source: Dorigan and Hushon, 1976
C-3
C6HSN02 or @-N02
123.11
S.6 - S.7·C
2I0.9·C at 760 torr
0.1 - 0.2 gm/100 ml at 20·C1.0 gm/100 ml at 100·C
ethanol, diethyl ether, acetone,benzene, lipids
0.284 mmHg at 2S·C600 mmHg at 200·C
4.24 (air = 1.0)
hexane/water - 3.18 at 24.4·C
1.199 gm/ml at 2S·C
87.8·C
482.2·C
1.682 cp at 30·C
10-4 mmoles/l
water systems are usually too low to measure (Pierce, 1979). Nitrobenzene
in water from an i ndustri a1 spi 11 is removed by treatment with acti vated
charcoal.
There are no data available on manmalian toxicity of nitrobenzene in
gested in drinking water.
Ingestion from Food
There are reports of nitrobenzene poisoning resulting from its uses as
false almond oil in baking, rubbing on the gums to ease toothache, contami
nation of alcoholic drinks, and contamination of food (Nabarro, 1948).
Leader (1932) reported a case of nitrobenzene poisoning in a child who was
given "oil of a1monds" for relief of a cold. Acute nitrobenzene poisoning
has also occurred from ingestion of denatured alcohol (Donovan, 1920; Wirt
schafter and Wo1paw, 1944). These cases are typical of accidental nitroben
zene ingestion. Nitrobenzene is not an approved food additive (Dorigan and
Hushon, 1976).
A bioconcentration factor (BCF) relates the concentration of a chemical
in aquatic animals to the concentration in the water in which they live.
The steady-state BCFs for a lipid-soluble compound in the tissues of various
aquatic animals seem to be proportional to the percent lipid in the tissue.
Thus, the per capita ingestion of a lipid-soluble chemical can be estimated
from the per capita consumption of fish and shellfish, the weighted average
percent lipids of consumed fish and shellfish, and a steady-state BCF for
the chemical.
Data from a recent survey on fish and shellfish consumption in the
United States were analyzed by SRI International (U.S. EPA, 1980). These
data were used to estimate that the per capita consumption of freshwater and
estuarine fish and shellfish in the United States is 6.5 g/day (Stephan,
C-4
1980). In addition, these data were used with data on the fat content of
the edible portion of the same species to estimate that the weighted average
percent 1i pi ds for consumed freshwater and estuari ne fi sh and she 11 fi sh is
3.0 percent.
No measured steady-state bioconcentration factor (BCF) is available for
nitrobenzene, but the eQuation IILog BCF = (0.85 Log P) - 0.70 11 can be used
(Veith et al., 1979) to estimate the BCF for aQuatic organisms that contain
about 7.6 percent lipids (Veith, 1980) from the octano1/water partition co
efficient (P). Based on an average measured log P value of 1.84 (Hansch and
Leo, 1979; Dec, et al., Manuscript), the steady-state bioconcentration fac
tor for nitrobenzene is estimated to be 7.31. An adjustment factor of
3.0/7.6 = 0.395 can be used to adjust the estimated BCF from the 7.6 percent
lipids on which the equation is based to the 3.0 percent lipids that is the
weighted average for consumed fish and shellfish. Thus, the weighted aver
age bioconcentration factor for nitrobenzene and the edible portion of all
aQuatic organisms consumed by Americans is calculated to be 7.31 x 0.395 =
2.89.
Inhalation
Nitrobenzene is readily absorbed through the lungs with retention of up
to 80 percent (Piotrowski, 1967). There are reports of nitrobenzene poison
ing from inhalation of an exterminator spray for bedbugs which was sprayed
on a child's mattress (Stevenson and Forbes, 1942; Nabarro, 1948). Poison
ings have also resulted from inhaled nitrobenzene used as a scent in perfume
and soap (Dorigan and Hushon, 1976). Chronic and acute poisonings from ex
posure to nitrobenzene vapor in production plants are well documented (Dori
gan and Hushon, 1976; Browning, 1950; Zeligs, 1929; Hamilton, 1919), but
since nitrObenzene is also absorbed through the skin, industrial poisoning
cannot be attributed to inhalation alone. A worker exposed to nitrobenzene
C-5
at 5 mg/m3, the current Occupational Safety and Health Administration
(OSHA) standard (40 CFR 1910.1000), would absorb 18 mg/day through the lungs
in 6 hours (Piotrowski, 1967).
Dermal
Nitrobenzene is highly fat-soluble and can be absorbed through the skin
at rates as high as 2 mg/cm2/hr (Dorigan and Hushon, 1976). Medical lit-
erature contains many reports of poisonings from absorption of nitrobenzene
in shoe dyes and laundry marking ink. These reports were common during the
19th century and the first half of this century.
There have been reports of cases of shoe dye poisoning in an army camp
(Levin, 1927), and in children who were given freshly dyed shoes (Zeitoun,
1959; Graves, 1928; Levin, 1927). The most frequent signs and symptoms were
dizziness, bluish color of lips and nails (cyanosis), headache, and some-
times coma.
Cyanosis and poisoning of newborns who came in contact with diapers or
pads containing marking ink were very common. Generally this occurred when
the diapers or pads were freshly stamped by the hospital laundry (Etteldorf,
1951; Ramsay and Harvey, 1959; MacMath and Apley, 1954; Zeligs, 1929; Ray
ner, 1886). Often the imprint of the ink could be seen on the infant's
skin. Removal of the diaper or pad and thorough washing of the skin usually
reduced toxic symptoms, although methylene blue and ascorbic acid have also
been used to relieve cyanosis. The toxicity is often more severe in prema
ture infants who are in an incubator and exposed to the vapor as well as to
the dye on the cloth (Etteldorf, 1951). Washing of the marked diapers or
pads before their use removes the hazard of absorpti on of ni trobenzene or
aniline from the ink.
C-6
In Egypt, "pure bitter almond oil" (a mixture of 2 to 10 percent nitro
benzene and 90 to 98 percent cottonseed oil) has been rubbed on babies to
remove crusts from the skin and to protect the children from other diseases.
Zeitoun (1959) reported cases of nitrobenzene poisoning seen in Alexandria
hospitals as a result of this practice.
Hamilton (1919) reported a case of chronic nitrobenzene poisoning in a
woman who used it as a cleaning fluid for many years. The continuous dermal
absorption caused her to experience symptoms of multiple neuritis, extreme
indigestion and hemorrhages of the larynx and pharynx.
Dermal absorption of nitrobenzene is the cause of many of the chronic
and acute toxic effects seen in nitrobenzene workers (inhalation also ac-
counts for industrial toxicity although the routes of exposure often cannot
be distinguished). The amo~nt of cutaneous absorption is a function of the
ambient concentration, the amount of clothing worn, and the relative humidi
ty (high humidity increases absorption) (Dorigan and Hushon, 1976). A
worker exposed to the current OSHA standard (40 CFR 1910.1000), 5 mg/m3,
could absorb up to 25 mg in six hours, and one-third of that amount would
pass through the skin of a clothed man (Piotrowski, 1967). Pacseri and
Magos (1958) measured ambient nitrobenzene in industri a1 pl ants and found
levels of up to eight times the current limit.
Hamilton (1919) reported a case of acute, fatal, nitrobenzene poisoning
that resulted from a soap factory worker spill ing "oil of mirbane ll on his
clothes. Immediate removal of the contaminated clothing would probably have
prevented his death.
There are reports of acute and chronic poisoning due to skin absorption
of dinitrobenzene by workers in munitions and nitrobenzene plants. Dinitro-
benzene is believed to be much more toxic than nitrobenzene (Malden, 1907).
C-7
Ishihara, et ale (1976) reported a case of poisoning where a worker handled
a cleaning mixture containing 0.5 percent dinitrobenzene. The worker wore
gloves, but the dinitrobenzene penetrated the gloves to cause acute symptoms
of methemoglobinemia and hemolytic jaundice. Rejsek (1947) described dini
trobenzene diffusion through the skin of munitions workers. Some of these
workers with chronic dinitrobenzene poisoning experienced an acute crisis
after exposure to sun or dri nk i ng a1coho1 (beer). A1coho 1 ingest i on or
chronic alcoholism can also lower the lethal or toxic dose of nitrobenzene
(Dorigan and Hushon, 1976). This acute reaction could occur as late as six
weeks after toxic symptoms disappeared.
A1though there are many 1i terature references deal i ng wi th occupat i ona1
exposure to nitrobenzene, there are few, if any, reports of nitrobenzene ex
posure resulting from water ... intake. Therefore, data derived from occupa
tional exposure will be used to develop information for establishing the
water quality criterion in this document.
PHARMACOKINETICS
Absorption
Nitrobenzene absorption can occur by all poss i b1e routes, but it takes
place mainly through the respiratory tract and skin. At 5 mg/m3, a nitro
benzene worker can absorb 18 mg through the lungs and 7 mg through the skin
in 6 hours (Piotrowski, 1967). On the average, 80 percent of the nitro
benzene vapor is retained in the human respiratory tract (Piotrowski, 1977).
Nitrobenzene, as liquid and vapor, will pass directly through the skin.
The rate of vapor absorption depends on the air concentration, ranging from
1 mg/hr at 5 mg/m3 concentration to 9 mg/hr at 20 mg/m3 • Air tempera
ture does not affect the absorption rate, but an increase of relative
humidity from 33 to 67 percent will increase the absorption rate by 40
C-8
percent. Work clothes reduce cutaneous absorption of nitrobenzene vapors by
20 percent (Piotrowski, 1977).
Maximal cutaneous absorption of liquid nitrobenzene is 0.2 to 3
mg/cm2/hr depending on skin temperature. Elevated skin temperature will
increase absorption. Absorption wi 11 decrease with duration of contact.
Cutaneous absorption can be significant in industry, since contamination of
the skin and clothing of dye manufacture workers may reach levels of 2 and
25 mg/cm2, respectively (Piotrowski, 1977).
Oistribution
Upon entry into the body, nitrobenzene enters the bloodstream, where it
reacts with the hemoglobin to form its oxidation product, methemoglobin.
Methemoglobin has a reduced affinity for oxygen, and the reduced oxygen car
rying capacity of the blood is the cause of most of the toxic effects of
nitrobenzene, including its lethality. Methemoglobin levels from nitroben
zene have ranged from 0.6 gm/100 ml in industrial chronic exposure to 10
gm/100 ml in acute poisoning (Pacseri and Magos, 1958; Myslak, et al. 1971).
The normal methemoglobin level is 0.5 gm/lOO ml. Under normal conditions
methemoglobin will slowly be reduced to oxyhemoglobin, the normal form of
blood hemoglobin.
Pacseri and Magos (1958) have demonstrated that sulfhemoglobin is also
formed in the blood after chronic exposure to nitrobenzene. In nitrobenzene
workers, they found average sulfhemoglobin levels of 0.27 gm/100 ml (com
pared to the upper limit of normal of 0.18 gm/100 ml). Pacseri postulated
that since blood sulfhemoglobin disappears more slowly than methemoglobin,
it is a more sensitive indicator of nitrobenzene exposure. Sulfhemoglobin
may be more specific than sensitive because methemoglobin is normally found
in the blood whereas sulfhemoglobin is not.
C-9
Ueh1eke (1964) measured the velocity of methemoglobin formation from ni
trobenzene in cats. He found the rate to be variable and not related to the
blood concentration of nitrobenzene, although the methemoglobin formation
velocity was maximal in each animal at the time of highest blood concentra
tion of nitrobenzene. He a1 so found that metabolites of nitrobenzene are
able to oxidize hemoglobin. Methemoglobin formation from nitrobenzene has
a1so been demonstrated in vi tro (Dori gan and Hushon, 1976, Kusumoto and
Nakajima, 1970).
Further indications of the presence of nitrobenzene in the blood are the
production of hemolytic anemia after acute exposure (Harrison, 1977) and the
alteration of the sodium and potassium permeability of erythrocytes by de
rivatives of nitrobenzene (Cooke, et al. 1968).
Nitrobenzene is very 1i~id soluble, with an oil to water partition coef
ficient of 800. In a rat study, the ratio of the concentration of nitroben
zene in adipose tissue versus blood in internal organs and muscle was ap
proximately 10:1 one hour after an intravenous administration (Piotrowski,
1977). Rabbits intubated with 0.25 ml of nitrobenzene had 50 percent of the
compound accumulated unchanged in tissues within two days after the intuba
tion (Dorigan and Hushon, 1976).
Dresbach and Chandler (1918) have shown cerebellar disturbances in dogs
and birds exposed to nitrobenzene vapor. A histologic study attributed
these effects to changes in the Purkinje cells of the cerebellum. Reports
of the effect of nitrobenzene on the 1iver vary from descri pt ion of 1i ver
damage from accumulated nitrobenzene (Dorigan and Hushon, 1976) to the
statement that nitrobenzene does not cause severe renal or 1i ver damage
(Goldstein, 1975). Goldwater (1947) has described hyperplasia of the ery
thropoietic centers of the bone marrow in workers chronically exposed to ni-
C-IO
trobenzene, but he concluded that the hyperplasia is a secondary result of
the hemolytic effect of the compound. Makotchenko and Akhmetov (1972) ob
served secretory changes of the adrenal cortex of guinea pigs given nitro
benzene every other day at a dose of 0.2 gm/kg for six months.
Metabolism
Available information on nitrobenzene metabolism is based on animal ex
periments and fragmentary human data. There are two main metabol ic path
ways: (l) reduct i on to an il i ne fo 11 owed hy hydroxyl at i on to ami nopheno1s
and (2) direct hydroxylation of nitrobenzene to form nitrophenols. Further
reduction of nitrophenols to aminophenols may also occur (Piotrowski, 1977).
The rate of nitrobenzene metabol ism is independent of the dose in later
stages of acute or chronic intoxication. This can cause its accumulation in
high-lipid tissues (Dorigan .nd Hushon, 1976).
The reduction of nitrobenzene to aniline occurs via the unstable inter
mediates, nitrosobenzene and phenyl hydroxylamine, both of which are toxic
and have pronounced methemoglobinemic capacity. The reactions occur in the
cytop1asmi c and mi crosoma1 fract ions of 1iver ce11 s by the nitro-reductase
enzyme system (Fouts and Brodie, 1957). This enzyme system is active in
mice, guinea pigs, and rabbits, and is less active in rats and dogs. The
aniline is then excreted as an acetyl derivative or hydroxylated and ex
creted as an aminophenol. Reddy, et ale (1976) showed that the gut flora of
rats was needed for the reduction of nitrobenzene and subsequent methemoglo
bin formation.
The hydroxylation of nitrobenzene to nitrophenols does not occur in the
microsomal fraction. The reaction proceeds via a peroxidase in the presence
of oxygen (Piotrowski, 1977).
Robinson, et a1. (1951) studied nitrobenzene metabolism in the rabbit
us i ng 14C_l abe1ed materi a1. The main metabo1ic product found was p-ami no-
C-ll
phenol (35 percent) which was formed via phenyl hydroxyl amine. Seven phenols
and aniline were detected as metabolites within 48 hours of a dose of 150 to
200 mg/kg body weight of nitrobenzene. Nitrobenzene was retained somewhat
in the rabbits; its metabolites were detected in urine one week after
dosing. Little unchanged nitrobenzene was excreted in the urine. The major
urinary metabolites were p-aminophenol, nitrophenols, and nitrocatechol.
These const ituted 55 percent of the uri nary metabo 1i tes and were excreted
conjugated with sulfuric and glucuronic acids. About 1 percent of the dose
was expired as radiolabeled carbon dioxide.
Yamada (1958) studied nitrobenzene metabolism in rabbits in a 3-month
subcutaneous exposure study. He found that urinary excretion of detoxifica
tion products varied in the early stage of exposure, but did not in the
later stages. The reduction~and hydroxylation pathways all became depressed
during the later stages of this chronic poisoning study.
Parke (1956) reports metabo1ites of nitrobenzene i so 1ated four to fi ve
days after administering 0.25 mg/kg orally as a single dose in the rabbit
(Table 2).
An investigation of the metabolism of 14C-nitrobenzene in the cattle
tick, Boophilus microplus, and spider, Nephia plumipes, was done by Holder
and Wilcox (1973). They found that the tick metabolized nitrobenzene to
nitrophenol and aniline whereas no free phenols were found as metabolites
inthe spider. Aniline was the major metabolic product in both species.
Nitrobenzene, if present in sufficiently small amounts in water, can be
degraded by some bacteria, such as Azobacter agilis. Nitrobenzene tends to
inhibit its own degradation at concentrations above 0.02 to 0.03 mg/l (Dor
igan and Hushon, 1976; Lu and Metcalf, 1975).
Lu and Metcalf (1975) studied nitrobenzene in a model aquatic ecosystem
to assess biodegradation and biomagnification. The ecosystem consisted of
C-12
TABLE 2
Metabolic Fate of a Single Oral Dose (0.25 g/kg) of [14C] Nitrobenzenein the Rabbit During 4-5 Days After Dosinga
Metabolite
Respiratory C02
Nitrobenzene
Aniline
Percentage of Dose (average)
2 in expired air
o-Nitrophenol
m-Nitrophenol
p-Nitrophenol
o-Aminophenol
m-Aminophenol
p-Aminophenol
4-Nitrocatechol
Nitroquinol
p-NitrophenylMercapturic acid
(Total urinary radioactivity)
Metabolized nitrobenzenein feces
Metabolized nitrobenzenein tissues
Total accounted for
0.1
9
9
3 60 total
4 58 in
31 urine
0.7
0.1
0.3
(58)
9**
15-20
85-9m¥l
aSource: Parke, 1956* 0.5% in the urine and 0.1% in the expired air.+ 0.3~ in the urine and 0.1% in the expired air.** 6% of the dose was present in the feces as p-aminophenol.
C-13
green filamentous algae, Oedogonium cardiacium, snails, Physa, water fleas,
Daphnia magna, mosquito larvae, Culex quinquifasciatus, and mosquito fish,
Gambusia affinis, under controlled atmospheric conditions. 14C-labeled
nitrobenzene 0.005 to 0.5 mg/m3 (0.01 to 0.1 ppm) was added to the water
and animals were removed for analysis after 24 to 48 hours. The radiolabel
ed metabo1i tes were extracted and separated by th i n 1ayer chromatography.
The distribution of nitrobenzene and its degradation products is listed in
Table 3.
Nitrobenzene was neither stored nor ecologicaly magnified, but was re
duced to aniline in all organisms, acetylated in fish and water extracts
only, and hydroxylated to nitrophenols by mosquito larvae and snails. The
metabolites of nitrobenzene formed by the different organisms are illustra
ted in Figure 1.
Excretion
In man, the primary known excretion products of nitrobenzene are p-ami
nophenol and p-nitrophenol which appear in the urine after chronic or acute
exposure. In experimental inhalation exposure to nitrobenzene, p-nitrophe-
nol was formed with the efficiency of 6 to 21 percent. The efficiency of
p-aminophenol formation is estimated .f:rom observation of acute poisoning
cases where the molar ratio of excreted p-nitrophenol to p-aminophenol is
two to one, since p-aminophenol is not formed at a detectable level in short
subacute exposure (Piotrowski, 1977).
Ikeda and Kita (1964) measured the urinary excretion of p-nitrophenol
and p-aminophenol in a patient admitted to a hospital with toxic symptoms
resulting from a 17-month chronic industrial exposure to nitrobenzene. The
results of their study are shown in Figure 2, which demonstrates that the
rate of excretion of the two metabolites parallels the level of methemoglo-
bin in blood. The authors exposed five adult rats to nitrobenzene vapor at
C-14
TABLE 3
Distribution of Nitrobenzene and Degradation Products in Model Aquatic Ecosystem*
Nitrobenzene equivalents. ppmRfa H2O Oedognoium Daphnia Culex Physa Gambusia
(alga) (daphnia) {mosquito} {snai l} {fish}
Total 14C 0.53755 0.0690 0.1812 0.5860 0.6807 4.9541
Nitrobenzene 0.72 0.50681 0.0162 0.0709 0.3952 0.3886 4.0088
Anil ine 0.60 0.01262 0.0032 0.0079 0.0272 0.0169 0.3527
()Aminophenolsb 0.20 0.00106 0.0080 0.0315 0.0986
I..... Nitrophenolsb 0.10 0.00466 0.0016 0.0394 0.1226 0.2190 0.0847U1
Polar 0.0 0.00896 0.0240 0.0315 0.0138 0.0393 0.1130
Unextractable 0.00164
*Source: Lu and Metcalf, 1975a TLC with benzene:acetone:Skellysolve B {bp 60-680 C}:diethylamine=65:25:25:5 {v/v}.b The isomers could not be separated reliably because of small amounts and similar Rf values.
-•-~
ozw:cGO_!ii:IE ..c~I ~Q,~....-"0'--•~-o 0z Ew =-.f!oa:I--ZIQ,
200
150
100
50
HOURS
FIGURE 1
Relative detoxication capacities of key organisms of a model aquaticecosystem following treatment with radioactive nitrobenzene.
Source: Lu and Metcalf, 1975
C-16
HOSPITAL DAYS
III.::l c:
,.. en 0III 0 -
'0 ~.;;;; .. EIII _ '0
.... 0 <,.5 10 15
OJClI....J:.
"enQ
•40
-i
0-i»r-
J:»ms:0C)r-0CD-
..... Z(Q
9.....-00 Qo3 s:~
mC7 -i0 :J:0 »Q. m
s:0Qr-0CD-Z.....x
6
8
4
2
o
16
14
10
12
255302520
P"·/;, ••" ..~I
-'--II
I12,,,,,,
.-_C0----.".,,,,,,,
.._4-• •••
15
.OS6
I
o
o...J
ozwJ:a.oz;:E .....
« ~I _
Q. ...:l
GO •
..... E0 ......... II)
Ql...J-o 0z Ew ~J: Ea. ....oa:::~
zI
Q.
AUGUST SEPTEMBER
FIGURE 2
Changes in the levels of total hemoglobin and methemoglobin in blood andof p-nitrophenol and p-aminophenol in urine. The usual daily volume ofurine was about 1 litre.
Source: Ikeda and Kita, 1964
C-17
125 mg/m3 {25 ppm} for eight hours and measured the subsequent excretion
of p-aminophenol and p-nitrophenol. The results are shown in Figure 3. The
urinary excretion ratio of p-aminophenol and p-nitrophenol corresponded to
their findings in the human case.
Studies of nitrobenzene concentrations in the blood of an acutely ex
posed person indicate that the compound remains in the human body for a pro
longed period of time. Similar observations have been made from excretion
of the two urinary metabol ites in patients treated for acute or subacute
poisoning. The excretion coefficient of urinary p-nitrophenol, followed for
three weeks, is about 0.008 per hour. Metabolic transformation and excre
tion of nitrobenzene in humans is slower by an order of magnitude than in
rats or rabbits {Piotrowski, 1977}.
Because of the slow rate of nitrobenzene metabolism by humans, the con
centration of p-nitrophenol in the urine increases for about four days
during exposure and the concentration on the first day is only about 40 per
cent of the peak value. An estimate of the mean daily dose of nitrobenzene
in chronic industrial exposure can be obtained by the measurement of urinary
p-nitrophenol in specimens taken on each of the last three days of the work
week. The level of nitrobenzene exposure can be approximated using the for
mula y = 0.18z, where y is the daily excretion of urinary p-nitrophenol in
mg/day and z is the mean daily dose of absorbed nitrobenzene in mg {Pio
trowski, 1967}. The extended systemic retention and slow excretion of meta
bo1i tes of nitrobenzene inman is determi ned by the low rate of metabo 1ic
transformation {reduction and hydroxylation} of the nitrobenzene itself.
The conjugation and excretion of the metabolites, p-nitrophenol and p-amino
phenol, is rapid {Piotrowski, 1977}.
The urinary metabolites in man account for only 20 to 30 percent of the
nit robenzene dose; the fate of the rest of the metabo1i tes is not known
c-18
100
,. Parent
o Nitrophenols
• Reduced (Aniline) m Aminophenois
~ Conjugated
80
0..,.
60-c:CD0~
CD -0a.
20
oOedogonium Daphnia Culex
FIGURE 3
Physa Gambusia
Excretion of p-nitrophenol and p-aminophenol in the urine of rats ex
posed to nitrobenzene.
Source: Ikeda and Kita, 1964
C-19
(Piotrowski, 1977). Parke (1956) studied 14C-nitrobenzene metabolism in
rabbits and was able to account for 8~ to 90 percent of the dose which was
administered by intubation. One percent of the nitrobenzene was exhaled as
CO2 in air, and 0.6 percent was exhaled as unchanged nitrobenzene. Fifty
eight percent of the dose appeared as urinary metabolites, p-aminopheno1,
nitropheno1s, aminopheno1s, nitrocatecho1s, and aniline. Thirty percent of
the nitrobenzene remained in the rabbit tissue four to five days after
dosing, and nine percent of the nitrobenzene metabolites were in the feces.
Urinary p-nitropheno1 in man is determined after hydrolysis of the con
jugated metabolites. Analytical methodology (of which there are several
methods) involves removal of interfering color substances, hydrolysis, ex
traction of p-nitropheno1, re-extraction into an aqueous system, reduction
to a p-aminopheno1, and reaction to indophenol, which is a blue colored pro
duct. The sensitivity is 5 ~g per sample (Piotrowski, 1977).
EFFECTS
Acute, Subacute, and Chronic Toxicity
Acute exposure to nitrobenzene can occur from accidental or suicidal in
gestion of the liquid nitrobenzene or ingestion as false bitter almond oil
in food or medicine. Cutaneous absorption causing acute toxic reactions can
result from wearlng wet, freshly dyed shoes (Levin, 1927); use of on diapers
or protective pads (Ette1dorf, 1951); use of soap or skin oil containing ni
trobenzene (Zeitoun, 1959); or from an untreated spill of nitrobenzene on
the skin in an industrial plant (Hamilton, 1919). The fatal dose of nitro
benzene in humans varies widely; values from less than 1 m1 to over 400 m1
have been reported (Wirtschafter and Wolpaw, 1944). Chronic toxic effects
in man generally result from industrial exposure to vapor that is absorbed
C-20
through the lungs or the skin. One case of chronic toxicity was reported in
a woman who used nitrobenzene as a cleaning solution for many years
(Hamilton, 1919).
Symptoms of chronic occupational nitrobenzene absorption are cyanosis,
methemoglobinemia, jaundice, anemia, sulfhemoglobinemia, presence of Heinz
bodies in the erythrocytes, dark colored urine, and the presence of nitro
benzene metabolites (e.g., nitrophenol) in the urine (Pacseri and Magos,
1958; Hamilton, 1919; Wuertz, et ale 1964; Browning, 1950; Malden, 1907;
Piotrowski, 1967).
The symptoms of dinitrobenzene poisoning include those found in nitro
benzene toxicity as well as abdominal pain, weakness, enlarged liver, and
basophilic granulations of red corpuscles (Beritic, 1956; Malden, 1907).
Oinitrobenzene poisoning also causes unequal responses in different exposed
workers.
The outstanding symptom of acute nitrobenzene poisoning is cyanosis as a
result of methemoglobin formation (up to 80 percent) (Piotrowski, 1967). If
the cyanosis is severe or prolonged the patient will go into coma and may
die. Often anemia is seen a week or two after acute poisoning as a result
of the hemolytic effect of nitrobenzene (Stevenson and Forbes, 1942). Sui-
cidal ingestion of nitrobenzene has been reported (Nabarro, 1948; Leinoff,
1936; Myslak, et ale 1971), and the compound has also been used unsuccess
fully to induce abortion (Nabarro, 1948; Dorigan and Hushon, 1976). Harri
son (1977) reported a case of poisoning from an aniline-nitrobenzene mixture
which was accidentally ingested from a pipette by a chemistry student. The
mortality due to ingested nitrobenzene in the above cases was variable, de
pending on the health of the patients and the treatments they received.
Common treatments include gavage, transfusions, oxygen therapy, methylene
blue, ascorbic acid, and toluidine blue. Treatment is usually directed to
C-21
reduce the methemoglobinemia which is the immediate effect, and often the
cause of death, in nitrobenzene poisoning. Death has resulted from intake
of less than one m1 of nitrobenzene (Wirtschafter and Wo1paw, 1944).
Some of the reported toxicity values are sunmarized in Table 4 (Fair
child, 1977). The term LDLo designates the lowest reported lethal dose
and TDLo is the lowest published toxic dose.
Levin (1927) demonstrated in~ production of methemoglobin by nitro
benzene in dogs, cats, and rats, but not in guinea pigs or rabbits. Dres
bach and Chandler (1918) found that nitrobenzene fumes caused cerebe11 ar
disturbances in dogs and birds, while blood changes were the principal toxic
effects in other mammals they studied. Reddy, et a1. (1976) reported a de
lay in methemoglobin formation in germ free rats by nitrobenzene and postu
lated that the gut flora of rats was responsible for the reduction of (~
vivo) and methemoglobin forming capacity of nitrobenzene. Sh imk in (1939)
measured the toxicity of nitrobenzene in mice when absorbed through the
skin. He found the minimum lethal dose to be 0.0004 m1/gm body weight by a
subcutaneous route of administration. The nitrobenzene caused respiratory
failure, reduction of the white blood cell count, and liver pathology in the
mice.
Yamada (1958) did a chronic toxicity study in rabbits that received a
subcutaneous dose of 840 mg/kg body wei ght per day for three months. He
found a decrease in erythrocyte number and hemoglobin content early in the
exposure. These values increased during the three months but did not return
to normal levels. Urinary excretion of detoxification products was variable
in the early stages of the exposure, but then all the detoxification reac
tions (reduction, hydroxylation, and acetylation) were depressed. As a
result of these Observations, Yamada divided this response in the rabbit
into three stages: initial response, resistance, and exhaustion.
C-22
The effects of subacute nitrobenzene exposure in rats were studied by
Kulinskaya (1974). Vasilenko and Zvezdai (1972) measured blood changes and
found sulfhemoglobin formation to be the most regular and persistant change
noted. Increased methemoglobin levels with Heinz body formation and anemia
were also seen.
The cytotoxicity of nitrobenzene to cultured Erl ich-Landschutz diploid
(ELD) cells was measured by Holmberg and Malmfors (1974). They found no
significant increase in cell injury after five hours incubation with nitro
benzene. However, a 3M nitrobenzene solution reduced cell proliferation by
50 percent in cultured hamster cells (Raleigh, et al. 1973). Oxygen con
sumption by cultured cells is increased by nitrobenzene (Biaglow and Jacob
son, 1977). Its derivatives are used to sensitize malignant cells .:!.!!. vitro
to radiation effects (Chapman, et al. 1974). The authors suggest that this
effect was due to radical oxidation and increased cellular damage.
Nitrobenzene derivatives have a wide variety of toxic effects. 1-Chloro
2,4-dinitrobenzene (ONCB) is a well known skin sensitizer in guinea pigs,
mice, and humans (Hamaguchi, et al. 1972; Jansen and Bleumink, 1970; Maurer,
et ale 1975; Weigand and Gaylor, 1974; Noonan and Halliday, 1978). Cooke,
et al. (1968) showed that nitrobenzene derivatives react with cell membranes
to alter sodium-potassium conductance, and sometimes affect action poten
tials of nerve cells.
m-Dinitrobenzene is a potent methemoglobin former, and is more toxic
than nitrobenzene (Ishihara, et al. 1976; Pankow, et al. 1975). Pentachlo
ronitrobenzene (PCNS) is a common fungicide with varying toxic effects in
different mammalian species (Courtney, et al. 1976).
Some of the toxic effects of nitrobenzene are summarized in Appendix A
(Dorigan and Hushon, 1976).
C-23
Animal
TABLE 4
Acute Toxicity Values*
Route Toxic Dose
Human (female) oral TOLO : 200 mg/kg
Human oral LOLo: 5 mg/kg
Rat oral LOSO: 640 mg/kg
Rat skin LOSO : 2,100 mg/kg
Rat i.p. LOSO: 640 mg/kg
Rat s.c. LOLo: 800 mg/kg
Mouse s.c. LOLO: 286 mg/kg
Dog oral LOLO: 750 mg/kg
Dog i.v. LOLO: 150 mg/kg
Cat oral LOLo: 2,000 mg/kg
Cat skin LOLo: 25 g/kg
Rabbit oral LOLo: 700 mg/kg
Rabbit skin LOLo: 600 mg/kg
Guinea pig i.p. LOLO: 500 mg/kg
*Source: Fairchild, 1977
Aquatic toxicity: TLm at 96 hours: 10-100 mg/l (ppm).
C-24
Synergism and/or Antagonism
Alcohol has a synergistic effect on nitrobenzene poisoning. Ingestion
of an alcoholic beverage induced immediate acute toxic symptoms, including
coma, in a worker who had apparently recovered from the effects of chronic
nitrobenzene exposure. A1coho 1 i nges t ion or chron icalcoho1ism can lower
the lethal or toxic dose of nitrobenzene (Dorigan and Hushon, 1976). In
subchronic dinitrobenzene poisoning, drinking of one beer or exposure to sun•
can bring on an acute crisis as late as six weeks after the disappearance of
other symptoms (Rejsek, 1947).
Smyth, et ale (1969) studied the synergistic action between nitrobenzene
and 27 other industrial chemicals by intubation in rats. Most of the com
pounds tested did not alter the LDSO • In another study, ingestion of 2 to
20 ml of ethanol increased the severity of reaction to a 0.1 ml intravenous
dose of nitrobenzene in rabbits. This observation agrees with the clinical
data on the synergism of ethanol and nitrobenzene (Dorigan and Hushon, 1976).
Kaplan, et ale (1974) studied the effect of caffeine, an inducer of mi-
crosomal enzymes, on methemoglobin formation by nitrobenzene in rats.
Methemoglobin was formed and then decreased in induced animals. The in-
creased microsomal enzyme level increased the rate of metabolism and excre-
tion of nitrobenzene and thus caused a rapid decline of methemoglobin levels.
Teratogenicity
There is a paucity of information on the teratogenic effects of nitro
benzene. In one study (Kazanina, 1968b), 125 mg/kg was administered subcu
taneously to pregnant rats during preimplantation and placentation periods.
Delay of embryogenesis, alteration of normal placentation, and abnormalities
in the fetuses were observed. Gross morphogenic defects were seen in four
of 30 fetuses examined.
C-2S
Changes in the tissues of the chorion and placenta of pregnant women who
used nitrobenzene in the production of a rubber catalyst were observed. No
mention was made of the effects on fetal development or viability (Dorigan
and Hushon, 1976). Menstrual disturbances after chronic nitrobenzene expo
sure have also been reported.
Garg, et al. (1976) tested substituted nitrobenzene derivatives for
their abil ity to inhibit pregnancy in albino rats. Two of the compounds
tested (p-methoxy and p-ethoxy derivatives) inhibited implantation 100 per
cent when administered on days one through seven after impregnation.
The available data, although sketchy, indicate that women who are or
wish to become pregnant should avoid exposure to nitrobenzene. Further
studies of nitrobenzene teratogenicity in mammals are needed.
Mutagenicity
Chiu, et al. (1978) tested nitrobenzene and 53 commercially available
heterocyclic and aliphatic nitro- compounds for mutagenicity using the Ames
Salmonella typhimur;um strains TA 98 and TA 100. They reported that 34 of
the 53 compounds tested were mutagenic. Nitrobenzene was not found to be
mutagenic.
Trinitrobenzene was mutagenic in two in vitro assays, the Ames
Salmonella microsomal assay and the mitotic recombination assay in yeast
(Simmon, et al. 1977). Other nitrobenzene derivatives have demonstrated mu
tagenicity in ~ vitro assays, so that the mutagenicity of nitrobenzene is
still in question and additional work is needed in this area.
Carcinogenicity
The available literature does not demonstrate the carcinogenicity of ni
trobenzene, however, some nitrobenzene derivatives have demonstrated carcin
ogenic capacities. For example, pentachloronitrobenzene (PCNS) has induced
hepatomas and papillomas in mice (Courtney, et al. 1976).
C-26
1-Fluoro-2,4-dinitrobenzene (DNFB) was demonstrated by Bock, et al.
(1969) to be a promoter of skin tumors in mice, although it does not induce
them when administered alone.
Carcinogenic activity is frequently a general characteristic of struc
turally related compounds (Arcos and Argus, 1974). Because of the struc
tural similarity of nitrobenzene to the above nitrobenzene derivatives, ni
trobenzene should be regarded as a suspect carcinogen. The same conclusion,
based on more circumspect reasoning, was reached by Dorigan and Hushon
(1976). This suspicion, while strong enough to warrant the testing of ni
trobenzene for carcinogenicity, is not sufficiently strong to reconmend a
criterion based on carcinogenicity.
C-27
CRITERION FORMULATION
Existing Guidelines and Standards
The maximum allowable concentration of nitrobenzene in air in industrial
plants is 5 mg/m3• This value was set by the joint ILO/WHO Committee on
Occupational Health in 1975 (Goldstein, 1975). The OSHA standard for nitro
benzene in air is 5 mg/m3 (1 ppm) set in 1977 (40 CFR 1910.1000). This is
also the limit in Germany and Sweden while the exposure limit in the USSR is
3 mg/m3 (Dorigan and Hushon, 1976).
There are no standards for nitrobenzene 1eve1sin water. Ni trobenzene
was not listed among the substances for which a maximum water concentration
has been set.
Current Levels of Exposure
Extrapolating from Piotrowski's exposure data, a worker exposed to the
current occupational standard of 5 mg/m3 (1 ppm) nitrobenzene for an
eight-hour work day would absorb approximately 24 mg by inhalation and 9 mg
cutaneously. The maximum eight-hour uptake would be 33 mg, which is less
than the "reasonable safe" level of 35 mg/day (Dorigan and Hushon, 1976).
Doses of up to 70 mg/day have been reported for factory workers and up to 80
mg/day have been reported in a dye stuff factory in England (Piotrowski,
1967) •
Nitrobenzene can be a contaminant in industrial wastewater, and compa
nies utilizing or producing nitrobenzene are required to monitor its level
in their effluent waste. Using gas chromatography the minimum detectable
level of nitrobenzene in drinking water is 0.7 ng (Austern, et al. 1975).
Nitrobenzene may be vented to the atmosphere. The vents are usually e
quipped with absorbers or scrubbers, but some nitrobenzene vapor can
escape. Atmospheric nitrobenzene levels outside a plant are not monitored
C-28
by industry. Since inner plant levels are below the standard of 5 mg/m3
(l ppm) and nitrobenzene vapor accumulates at the floor level due to its
high density, the external air nitrobenzene concentrations are expected to
be very low (Dorigan and Hushon, 1976).
Special Groups at Risk
Workers in plants producing or using nitrobenzene have the greatest risk
of toxic exposure. At the current OSHA standard of 5 mg/m3 (1 ppm), a
worker could absorb as much as 33 mg/day. This is enough to produce symp
toms of chronic toxicity in some susceptible individuals (Dorigan and
Hushon, 1976). The amount of nitrobenzene absorbed by a worker via inhala
tion and cutaneous absorption can be estimated from the level of total (free
and conjugated) p-nitrophenol in urine as described by Piotrowski (1977).
Due to the current widespread use of disposable diapers and underpads in
hospitals, nitrobenzene poisoning in infants from laundry marking dyes, in
most cases, has been studied and corrected.
Pregnant women may be especially at risk with respect to nitrobenzene
due to transplacental passage of the agent. Individuals with glucose-6
phosphate dehydrogenase deficiency may also be at special risk (Calabrese,
et al. 1977; Djerassi, et al. 1975). Additionally, because alcohol ingestion
or chronic alcoholism can lower the lethal or toxic dose of nitrobenzene
(Rejsek, 1947; von Oettingen, 1941), individuals consuming alcoholic bever
ages may be at increased risk.
Basis and Derivation of Criterion
There are no established standards for nitrobenzene in water. Because
there are little or no data available on the toxicity of nitrobenzene in-
gested in drinking water, or on the teratogenic, mutagenic, or carcinogenic
effects of nitrobenzene in general, experimental testing is necessary before
C-29
a criterion can be derived from oral ingestion data. It is recommended that
testing in these areas of toxicity be implemented so that the effects of ni
trobenzene on mammals may be better understood.
Until more toxicological data on oral ingestion in animals are gener
ated, criterion levels must be estimated from occupational exposure data and
from organoleptic data. As reported, nitrobenzene produces a detectable
odor in water at a threshold (lowest discernible concentration) of 30 lJgll
(Austern, et ale 1975; U.S. EPA, 1970; Alekseeva, 1964). It should be
noted, however, that this criterion level is based on aesthetics rather than
health effects.
A water quality criterion (wQe) can be derived from the Threshold Limit
Value (TLV) of 5 mg/m3• This can be done by estimating the total daily
dose allowed by the TLV from both inhalation and dermal exposure. An inha
lation absorption coefficient of 0.8 will be used based on data provided by
Piotrowski (1967, 1977). Assuming an air intake of 10 m3/work day, the
portion of allowable dose by inhalation is 40 mg (5 mg/m3 x 10 m3/work
day x 0.8). The portion of the allowable dose by dermal exposure can be
calculated from the 7:18 ratio of dermal:inhalation exposure estimated by
Piotrowski (1967, 1977), i.e., 7/18 x 40 mg/work day = 16 mg/work day. Thus
the total allowable dose per work day is 56 mg (40 mg + 16 mg). The allow
able daily intake (AD!) can be calculated by adjusting for a 5/7 day work
week, i.e., 56 mg/work day x 517 = 40 mg/day.
Assuming 100 percent gastrointestinal absorption of nitrobenzene, a
daily water consumption of 2 liters, a daily fish consumption of 0.0065 kg,
and a bioconcentration factor of 2.89, the water quality criterion is:
40 mg = 19.8 mg/l2 liters + (2.89 x 0.0065)
C-30
Since the WOC using the TLV is well above the detectable odor level of
nitrobenzene, water containing this concentration of nitrobenzene would not
be aesthetically acceptable for drinking. Even though the limitations of
using organoleptic data as a basis for establishing a WOC are recognized, it
is recol1lnended that a WOC of 30 lJgll be established at the present time.
This level may be altered as more data are developed upon which to calculate
a WOC.
The analysis and recommendations generated in this document are based on
the literature available to date. If future reports indicate that nitroben
zene may be carcinogenic, mutagenic or teratogenic, a reassessment of the
WOC will be necessary.
C-31
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C-45
APPENDIX
Toxicological Effects of Nitrobenzene
Human
Route
Inhalation
Exposure Exposure Time
8 hrs./day for 17mos. factory worker
Response
Cyanosis, headache, fatigue methemoglobinemia (Ikeda and Kita,1964) •
Inhalation Poor ventilation 8 hrs./day for 1.5mos. factory workerpaint firm
8 hrs./day for 4.5mos.
Cyanosis, headache, fatigue, metheglobinemia, liver damage, hypotension (Ikeda and Kita, 1964).
Above plus: liver and spleen enlarged and tender, hyperalgesiain extremeties (Ikeda and Kita,1964) •
Inhalation
Inhalation
Inhalation
Inhalation
O. 2-0 •5 mg /l(40-100 ppm)
0.129 mg/m3
IILarge ll amountspoor ventilation
Acute
ca. 6 hours. Slight effects, e.g., headache,fatigue (von Oettingen, 1941).
Threshold level for electroencephalograph disturbance(Andreeshcheva, 1964).
Hospitalized:Day 1 - fatigue, headache, asthma
2 - vertigo, coma, cyanosis3 - labored breathing, urine
with almond odor7 - methemoglobinemia recovery
after 1 mo. (Ravault,et a1. 1946).
Burning throat, nausea, vomiting,gastrointestinal disturbances,cold skin, livid face, cyanosis(von Oettingen, 1941).
Organism
Human
Route
Inhalation
Inhalation
Inhalation
Inhalation
Inhalation
Inhalation
Exposure
6-30 lJgll
Acute
APPENDIX (Continued)
Exposure Time
Nitrobenzene factory worker
6 hrs.
Factory worker (rubber accelerator)
Factory worker(glass, porcelain)
Industrial exposure
Factory worker(filled containerswith nitrobenzene)
Response
Intermittent symptoms: cyanosis,pallor and jaundice, pharyngealcongestion, headache, changes inblood cell composition (increasedpolynuclears and eosinophils (vonOettingen, 1941).
Retained 80% of vapor in lungs,urinary excretion of p-nitrophenol(maximum in 2 hrs., still detectedafter 100 hrs.) (Salmowa, et ale1963).
Pregnant women: thickening of tissue in blood vessels, decreasedplacental absorption, necrosis inplacental tissue (Ferster, 1970).
Changes in bone marrow, increasedlymphoid cell production, impairment of copper metabolism and certain iron-eontaining enzymes(Yordanova, et ale 1971).
Disturbance of motor impulses(Zenk, 1970).
14 days: cyanosis, headache, backache, stomach ache, vomitingca. 21 days: drank- beer and fellunconscious, cyanosis, dilated pupils, retarded respiration, weakpulse1 yr.: intelligence dimmed2 yrs.: emaciated, atrophied
muscles
Organism
Human
Route Exposure
APPENDIX (Continued)
Exposure Time Response
3 yrs.: memory failed6 yrs: loss of perception of time
and space (Korsakoff1s syndrome) (Chandler, 1919).
()I
01>000
Rabbit
Cutaneousabsorption
Cutaneousabsorption
Cutaneousabsorption
Oral
Oral
Oral
Subcutaneousinjection
Dye used indiaper stamps
Shoe dye
0.5% byweight inpaper
333 ml
0.4 ml
0.8 mg/kg
ca. 7 hrs.
(Handled carbonpaper
From human milk
Single
Single
Daily for 3 mo.
Babies: cyanosis, rapid pulse,shallow respiration, vomiting,convulsions, recovery in 24 hrs.(von Oettingen, 1941).
Unconsciousness after consumptionof alcoholic beverages, death(Chandler, 1919).
Dermatitis(Calan and Connor, 1972)
Nurselings became cyanotic, recovery in 24 hrs. (mothers ate almond cake artificially flavoredwith nitrobenzene)(Dollinger, 1949).
Maximum dose with recovery reported following severe symptoms(von Oettingen, 1941).
Minimum lethal dose reported(von Oettingen, 1941).
Maximum dose not causing death(Yamada, 1958).
APPENDIX (Continued)
Organism Route Exposure Exposure Time--Rabbit Subcutaneous 10-14 mg/kg Single
injection
Cutaneous 700 mg/kg Singleabsorption
Intraperitoneal 500 mg/kg Singleinject ion
Intravenous 100 mg Daily or every 5days
()I Oral 9 gm 4 doses t one every..,.~ 15 minutes
Oral 4.8 gm Single
Oral 700 mg/kg Single
Oral 600 mg Single
Response
Minimum dose producing observableeffects; slow and lasting methemoglobinemia (von Oettingen t 1941)
After 52 hrs.: lethal(von Oettingen t 1941)
Reduced blood pressure and myocardial glycogen level(labunski t 1972).
Simultaneous doses of 2-20 m1 ethanol increased severity of poisoning (Matsumara and Yoshida t 1959).
Convu1sions t death (von Oettingen t1941; Chand1er t 1919).
lethal instantly (von Oettingen t1941; Chand1er t 1919).
lethal dose (Stecher t 1968).
Dizziness t loss of ref1exes t methemog10binemia t congestion of braintissue - 12 hrs. - death(Chandler. 1919).
Oral
Oral
300 mg
50 mg/kg
Single
Single
Fatigue for 1 week (Parke. 1956).
Tissue degenerationt especiallyheartt 1iver t kidney (Papageorgiouand Argoude1is t 1973)
APPENDIX (Continued)
Organism Route Exposure Exposure Time Response
Rabbit Oral 1 mg/kg Single Lowered hemoglobin, erthyrocytesand lymphocytes; increased leuco-cytes (Kazakova, 1956).
Oral 0.1 mg/kg Single Threshold toxic dose(Kazakova, 1956)
Guinea Inhalation Saturated air 2-5 hrs. Death following tremors, paralysispig (0.04 vol. %) of hind legs (Chandler, 1919).
Subcutaneous 0.2 gm/kg Every other day Hemolytic anemia, loss of weight,for 6 mos. decreased motor activity, fluxes
in urinary excretion of 17-hydroxy-0
corticosteroids (Porter-SilberI chromogens)
lJl(Makotchenko and Akhmetov, 1972).0
Oral ca. 3 gm Single 0.5 hrs: tremors, faint heartbeats,labored respiration
2 hrs: death (Chandler, 1919) .
Oral ca. 1. 2 gm Single Irrmediately motionless, then com-plete recovery (Chandler, 1919).
Oral 50 mg/kg 1 year Tissue degeneration, especiallyheart, 1i ver, kidney(Kazakova, 1956).
Oral 1 mg/kg Single Lowered hemoglobin, erythrocytes,lymphocytes; increased leucocytes(Kazakova, 1956).
Oral 0.1 mg/kg Single Threshold toxic dose (Kazakova,1956).
APPENDIX (Continued)
Organism Route Exposure Exposure Time--Rat Inhalation 5 mg/m3 8 hrs.
Inhalation ca. 0.03 mg/m3 Daily, up to98 days
Inhalation 0.06-0.1 mg/m3 70-82 days
Inhalation 0.008 mg/m3 73 days
Oral 600 mg/kg Single
() Intraperitonea1 800 mg/kg SingleI injectionU1
I-'
Subcutaneous 640 mg/kg Singleinjection
Subcutaneous 300 mg/kg Singleinjection
Subcutaneous 200 mg/kg Singleinjection or
100 mg/kg Daily for 10 days
Subcutaneous 125 mg/kg Singleinjection
Response
Metabolites excreted in 3 days(Ikeda and Kita, 1964).
Increased ability to form sulfhemoglobin in preference to methemoglobin (Andreeshcheva, 1970).
Cerebellar disturbances, inflamedinternal organs (Khanin, 1969).
No effect (Andreeshcheva, 1964).
LD50 (Smyth, et a1. 1969).
Lethal (Magos and Sziza, 1958).
Blood catalase activity decreasedcontinuously over 96 hrs.(Goldstein and Popovici, 1959).
LD (14 days) - methemoglobinemia,anemia, sulfhemoglobinemia(Brown, et ale 1975).
Methemoglobinemia, sulfhemoglobinemia, anemia(Zvezdai, 1972).
Delayed embryogenesis, abnormalfetal development and embryo deathchanges in polysaccharide composition of placenta(Kazanina, 1967, 1968a,c).
()I
U1N
Organism
Rat
Mouse
Cat
Route
Subcutaneousinjection
Cutaneousabsorption
Intraperitonealinjection
Intraperitonea1
Intraperitonealinjection
Intraperitonea1
injection
Inhalation
Inhalation
Oral
Exposure
100-200 mg/kg
480 mg/kg
1.23 gm/kg
1 gm/kg
20 mg/kg
12.3 mg/kg
Saturated air(0.04 vol. %)
2.4 gm
APPENDIX (Continued)
Exposure Time
Single
Single
Single
Single
Single
2-5 hrs.
2-3 hrs.
Single
Response
Su lfhemoglobin (most regu 1ar andpersistent form of hemoglobin) nitroxyhemoglobin. increased methemoglobin (Vasilenko and Zvezdai.1972) •
30 min: prostrate. motionless24 hrs: death (von Oettingen. 1941)
40 min.: 67% dead(Smith. et a1. 1967).
10-15 min: incoordination. comatoseshallow respiration
Several hrs.: regained coordinationImmediately before death: lostcoordination again. respiratoryarrest
48 hrs: death (Smith. et a1. 1967).
Lethal dose(Brown. et a1. 1975).
10 min.: 4.2% methemoglobinformed(Smith. et a1. 1967).
Death following tremors. paralysisof hind legs (Chandler. 1919).
Death
Death in 12-24 hrs. (von Oettingen.1941; Chandler. 1919).
()I
VIW
Organism
Dog
Route
Inhalation
Intravenousinjection
Oral
Oral
Oral
Exposure
IIThick vapor ll
150-250 mg/kg
28.8 gm plus6 gm
24 gm
2.4 gm
APPENDIX (Continued)
Exposure Time
1.5 hrs.
Single
2 doses, 0.5 hrs.apart
Single
Single
Response
Complete anesthesia and sleep(Chandler, 1919).
Minimum lethal dose - lowered bloodpressure, pulse rate increasedthen decreased; respiration stimulated until paralyzed(von Oettingen, 1941).
Immediate: agitation, then motionless
1 hr.: convulsions, then motionless4.5 hrs.: tremors, hind legs para
lyzed18 hrs.: death (Chandler, 1919).
Few hrs.: "stupid ll
12 hrs.: deep coma, slow respiration, lowered skin temperature,stomach strongly alkaline(Chandler, 1919).
1 hr: vomiting, then sleep continu-ing for 6 hrs.
6 hrs: appeared normal15-68 hrs: rigid muscles104 hrs: death (Chandler, 1919).
Oral
Oral
750-1000 mg/kg
500-700 mg/kg
Single
Single
Minimum lethal dose(von Oettingen, 1941).
Salivation, unrest, dizziness, tremors, increased pulse rate, sometimes convulsions (Chandler, 1919)
nI
U1~
.,!=!"
~l"l
51ii:l"lZ"I
'"~"I!Z
'"0..,..,r;l"l
~
'"'"0
Organism
Dog
Chicken
Pigeon
Route
Oral
Oral
Oral
Inhalation
Exposure
1.2 gm
2.4 gm
APPENDIX (Continued)
Exposure Time
Dai ly
Single
Single
1 hr.2-3 hrs.
Response
Formed methemoglobin continuouslyat "certain" concentration(Hashimoto t 1958).
Unsteady gait t recovery(Chandler t 1919).
Immediately unconscious12 hrs.: death (Chandler t 1919).
No effectsDeath (Chandler t 1919).